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Effects of DC loaded α-GalCer combined with tumor specific cytotoxic T lymphocytes on the growth of transplanted Heps hepatoma in mice
Wang Peng, Luan Zhiyong, Liu Junquan, Hang Min， Pan Jin, Zhang Nanzheng
2016, 43 (11):
806-811.
doi: 10.3760/cma.j.issn.1673-422X.2016.11.002
ObjectiveTo investigate the effects of dendritic cells (DCs) loading alphaGalactosylceramide (αGalCer) combined with tumor specific cytotoxic T lymphocytes (CTLs) on the growth of transplanted Heps hepatoma in mice. MethodsWe induced the augmentation of the DC cells and T lymphocyte derived from the mice bone marrow, and enabled them to be specific CTLs. DC cells loaded αGalCer in vitro. First we established a Heps liver cancer xenograft model, then divided the model mice into 4 groups by random number table method (n=9): control group (intravenous injection with physiological saline), CTL group, DC loading αGalcer group and DC loading αGalcer combined with CTLs group. After 2 weeks of intervention, we extracted the tumor tissue, weighed the tumor and calculated the inhibition rate of tumor. The expressions of Bax/Bcl2 cells in groups of transplanted tumor tissues were detected using immunohistochemistry and Western blotting. ResultsThe average tumor weight of CTL group, DC loading αGalCer group and combined treatment group were (1.07±0.15)g, (1.11±0.17)g, (0.79±0.14)g, respectively. All of them were lower than that of control group (1.69±0.23)g, with significant differences (t=14.176, P=0.023; t=12.351, P=0.034; t=18.672, P=0.000). The average tumor weight of combined treatment group was lower than those of the CTL group and DC loading αGalCer group, with significant differences (t=15.236, P=0.012; t=11.176, P=0.037). Compared to the CTL group (36.69%) and DC loading αGalCer group (34.32%), the combined treatment group had a higher tumor inhibition rate (53.25%; P=0.034, P=0.021). Immunohistochemical assay showed that the numbers of Baxpositive cells in CTL group, DC loading αGalCer group and combined treatment group were 35.83±0.75, 33.67±0.82, 41.17±1.17 respectively, and compared with the control group (21.67±2.16), the differences were statistically significant (t=-13.789, P=0.002; t=-15.116, P=0.001; t=-17.452, P=0.000). The numbers of Baxpositive cells in combined treatment group were different with CTL group and DC loading αGalCer group (t=-7.730, P=0.009; t=-5.872, P=0.011). The numbers of Bcl2positive cells in CTL group, DC loading αGalCer group and combined treatment group were 30.83±0.75, 31.67±1.03, 25.00±0.89, and compared with the control group (38.67±1.21), the differences were statistically significant (t=9.234, P=0.007; t=11.738, P=0.003; t=20.608, P=0.000). The numbers of Bcl2positive cells in combined treatment group were different with CTL group and DC loading αGalCer group (t=11.952, P=0.003; t=12.223, P=0.002). Western blotting test results showed that the expression levels of Bax in CTL group, DC loading αGalCer group and combined treatment group were 0.46±0.01, 0.42±0.03, 0.55±0.01, and compared with the control group (0.31±0.02), the differences were statistically significant (t=1.035, P=0.032; t=1.124, P=0.027; t=1.425, P=0.010). The expression level of Bax in combined treatment group was different with CTL group and DC loading αGalCer group (t=1.305, P=0.013; t=1.421, P=0.010). The positive expressions of Bcl2 in CTL group, DC loading αGalCer group and combined treatment group were 0.34±0.03, 0.33±0.02, 0.24±0.01, and compared with the control group (0.46±0.01), the differences were statistically significant (t=-1.123, P=0.025; t=-1.061, P=0.031; t=1.278, P=0.014); the positive expression level of Bcl2 in combined treatment group was different with CTL group and DC loading αGalCer group (t=1.160, P=0.021; t=1.219, P=0.015). ConclusionIt has synergistic killing effect on transplanted Heps hepatoma in mice using DC loading αGalCer combined with the tumor specific CTL.
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